Human Plasminogen Variant Chicago III

Summary

A new abnormal, variant, plasminogen Chicago III has been isolated from a patient with recurring deep vein thrombosis. Studies on the plasma fibrinolytic system of four family members showed no inheritance pattern. Kinetics of activation parameters of Chicago III plasminogen with different activators showed lowered catalytic rate constants from 159fold with urokinase, to 3fold with light (B) chain ‧ streptokinase complex, to 1.3fold with streptokinase. The Michaelis constants of activation of Chicago III plasminogen were 16fold higher with streptokinase, 6fold higher with light (B) chain ‧ streptokinase complex, and similar with urokinase. Each of the three activators exhibited different interaction characteristics with this variant zymogen, as they did with variant Chicago I and Chicago II plasminogens (previously reported). However, the latter variants were characterized by having normal catalytic rate constants of activation, with higher than normal apparent Michaelis constants of activation. Chicago III plasminogen, as well as Chicago I and II plasminogens, has a homogeneous population of molecules; the interpretation of the kinetic data was possible only in terms of a single population of molecules. The plasmins derived from Chicago I plasminogen, and also Chicago II and Chicago III plasminogens, have 100% active sites with normal amidase parameters. Basically, a single population of normal isoelectric forms were found in the Chicago III plasma, with three minor forms, about 11% of the total.

The kinetic parameters have permitted us to classify the four known plasminogen variants into three different classes. The Class A homozygote (Tochigi) has both an active center defect and a charge mutation difference with normal cleavage of the Arg₅₆₀-Val peptide bond; the Class B homozygote (Chicago I and Chicago II) has a K_m of activation defect with impaired activator binding and normal cleavage of the Arg₅₆₀-Val peptide bond; and, the Class C homozygote (Chicago III) has both a K_m of activation defect with impaired activator binding and a kcat of activation defect, and impaired Arg₅₆₀-Val peptide bond cleavage.

Keywords

Fibrinolysis - Deep vein thrombosis - Abnormal plasminogens - Variant plasminogen Chicago III - Plasminogen kinetics of activation parameters - Urokinase - Streptokinase - Light(B) chain ‧ streptokinase complex - Plasma plasminogen isoelectric forms

• References

• 1 Aoki N, Moroi M, Sakata Y, Yoshida N, Matsuda M. Abnormal plasminogen. A hereditary molecular abnormality found in a patient with recurrent thrombosis. J Clin Invest 1978; 61: 1186-1195
• 2 Wohl RC, Summaria L, Robbins KC. Physiological activation of the human fibrinolytic system. Isolation and characterization of human plasminogen variants, Chicago I and II. J Biol Chem 1979; 254: 9063-9069
• 3 Kazama M, Tahara C, Suzuki Z, Gohchi K, Abe T. Abnormal plasminogen, a case of recurrent thrombosis. Thromb Res 1981; 21: 517-522
• 4 Sakata Y, Aoki N. Molecular abnormality of plasminogen. J Biol Chem 1980; 255: 5442-5447
• 5 Aoki N. Genetic abnormalities of the fibrinolytic system. In: Menaché D, Surgenor D, Mac N, Anderson H. (eds) Hemophilia and Hemostasis. A R Liss; New York: 1981
• 6 Summaria L, Spitz F, Arzadon L, Boreisha IG, Robbins KC. Isolation and characterization of the affinity chromatography forms of human Glu- and Lys-plasminogens and plasmins. J Biol Chem 1976; 251: 3693-3699
• 7 Wohl RC, Summaria L, Arzadon L, Robbins KC. Steady state kinetics of activation of human and bovine plasminogens by streptokinase and its equimolar complexes with various activated forms of human plasminogen. J Biol Chem 1978; 253: 1402-1407
• 8 Wohl RC, Summaria L, Robbins KC. Kinetics of activation of human plasminogen by different activator species at pH 7.4 and 37° C. J Biol Chem 1980; 255: 2005-2013
• 9 Summaria L, Robbins KC. Isolation of a human plasmin-derived, functionally active, light (B) chain capable of forming with streptokinase and equimolar light (B) chain·streptokinase complex with plasminogen activator activity. J Biol Chem 1976; 251: 5810-5813
• 10 Robbins KC, Summaria L. An immunochemical study of human plasminogen and plasmin. Immunochem 1966; 3: 29-40
• 11 Hobart MJ. Genetic polymorphism of human plasminogen. Ann Hum Genet 1979; 42: 419-423
• 12 Raum D, Marcus D, Alper CA. Genetic polymorphism of human plasminogen. Am J Hum Genet 1980; 32: 681-689
• 13 Summaria L, Arzadon L, Bemabe P, Robbins KC, Barlow GH. Characterization of the NH₂-terminal glutamic acid and NH₂-terminal lysine forms of human plasminogen isolated by affinity chromatography and isoelectric focusing methods. J Biol Chem 1973; 248: 2984-2991
• 14 Hirsh J. Blood tests for the diagnosis of venous and arterial thrombosis. Blood 1981; 57: 1-8
• 15 Summaria L, Wohl RC, Boreisha IG, Robbins KC. A virgin enzyme derived from human plasminogen. Specific cleavage of the Arg₅₆₀-Val peptide bond in the DIP-virgin enzyme by plasminogen activators. Biochem 1982; 21: 2056-2059
• 16 Cederholm-Williams SA, De Cock F, Collen D. Kinetics of the reactions between streptokinase, plasmin and α₂-plasmin inhibitor. Eur J Biochem 1979; 100: 125-132
• 17 Bode W, Schwager P, Huber R. The transition of bovine trypsinogen to a trypsin-like state upon strong ligand binding. The refined crystal structures of the bovine trypsinogen-pancreatic trypsin inhibitor complex and of its ternary complex with Ileu-Val at 1.9 Ã… resolution. J Mol Biol 1978; 118: 99-112
• 18 Wohl RC, Sinio L, Robbins KC. Methods for Studying Fibrinolytic Pathway Components in Human Plasma. Thromb Res. 1982 27. in press